Functional characterization of LkERF-B2 for improved salt tolerance ability in Arabidopsis thaliana

The ethylene response factors have been reported to play critical roles in developmental and environmental responses in plants. In the present study, an ERF transcription factor gene was aimed to be identified from Larix kaempferi. Molecular characteristics and function of this gene were further explored. The result showed that a 1344 bp ERF transcription factor gene containing initiation and termination codon was obtained by RT-PCR and named LkERF-B2. LkERF-B2 gene encoded 447 amino acids containing a typical AP2/ERF domain. Alignment of predicted amino acid sequence of LkERF-B2 in various plant species showed that this ERF transcription factor was highly homologous (79.0%) with that of Picea sitchensi. To elucidate the function of LkERF-B2, LkERF-B2 overexpression vector was successfully constructed and transformed to Arabidopsis thaliana via dip flower. Compared with control plant, LkERF-B2 overexpressed transgenic A. thaliana showed a significantly higher survival rate under cold, heat, NaCl and drought stresses. NaCl stress analysis revealed that control and transgenic Arabidopsis were both flowering earlier under 100 and 150 mM/L NaCl treatment. While under 200-300 mM/L NaCl treatment, the growth of control plant was significantly inhibited compared with transgenic A. thaliana. Salt injury rate and salt injury index of transgenic Arabidopsis were lower than those of the control. Further investigation showed that transgenic Arabidopsis exhibited much higher content of chloroplast pigments under different NaCl concentration. Meanwhile, the activity of SOD and POD was also enhanced in transgenic A. thaliana. These results suggested that LkERF-B2 was a key transcription factor and could lead to enhanced salt stress tolerance.